An electrochemical aptasensor for the detection of bisphenol A based on triple signal amplification assisted by gold nanoparticles, hemin/G-quadruplex DNAzyme, and exonuclease I

A triple signal amplified electrochemical aptasensor for the detection of bisphenol A (BPA) was developed for the first time based on gold nanoparticles (AuNPs), hemin/G-quadruplex DNAzyme, and exonuclease I (Exo I) assisted amplification strategies. The BPA aptamer (Apt) hybridized with the capture probe (CP) was fixed on the gold electrode (GE) to form the double-stranded DNA (dsDNA) structure. When BPA was present, the Apt was detached from the GE surface by specific recognition between the BPA and Apt, forming BPA-Apt complexes in solution. The complexes could be selectively digested by Exo I, releasing BPA to participate in the cycle for binding to other Apt in dsDNA. The hybridization of the CP and auxiliary DNA (aDNA) within the detect probe DNA (dpDNA)-AuNP-aDNA nanocomplex allowed the nanocomplex to connect to the GE surface. The dpDNA interacted with K⁺ and hemin to produce hemin/G-quadruplex DNAzyme, which catalyzed H₂O₂ reduction, accelerated methylene blue (MB) oxidization, and further amplified the electrochemical signal. The integration of triple signal amplification strategies with aptamer-specific recognition enabled sensitive and specific detection of BPA. Under optimized conditions, the aptasensor exhibited a linear range of 0.1 pM–10 nM, with a low detection limit of 76 fM. Moreover, the designed aptasensor was successfully applied to detect BPA in lake water, fruit juice, and honey samples.

Graphical Abstract